FITC-β-Ala-Amyloid β-Protein (1-42)

β-Amyloid Aggregation Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs of your downstream experimental schemes).
Monomerization (HFIP-treated):
1. Solid Aβ peptide is dissolved in cold hexafluoro-2-propanol (HFIP). The peptide is incubated at room temperature for at least 1 h to establish monomerization and randomization of structure.
2. Conduct vacuum drying to remove all of HFIP, and the resulting peptide is stored as a film at -20 or -80 °C.
Oligomer preparation
3. The resulting film is dissolved in anhydrous DMSO at 5 mM and then dilutes into the appropriate concentration and buffer (cold PBS or serum-free and phenol red-free DMEM/F12 medium) with vortexing.
4. Next, the solution is age 24-48 h at 4-8 °C. The sample is then centrifuged at 14000g for 10 min at 4-8 °C; the soluble oligomers are in the supernatant. The supernatant is diluted 10-200-fold for experiments.
Fiber preparation:
3. The resulting film is dissolved in anhydrous DMSO at 5 mM and then dilutes into the appropriate concentration and buffer (10 mM HCl) with vortexing.
4. Next, the solution is age 24-48 h at 37 °C to obtain Aβ42 fibrils.
Note:
1) When performing step 1, if there is a small amount of insoluble matter, the HFIP treatment time can be extended (such as overnight incubation), vortexing, or short-term ultrasound can be used; if there is still a small amount of insoluble matter, it is recommended to remove it by centrifugation or filtration.
2) The aggregation form is unstable in the solution, it is recommended to prepare and use it immediately. If storage is required, it is recommended to store the peptides in a film form at -20 or -80 °C.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.